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Development and implementation of new approaches for proteomic characterization of bone tissues in oral surgery
Michalus, Iva ; Hynek, Radovan (advisor) ; Šebela, Marek (referee) ; Hrabal, Richard (referee)
1 Abstract Proteomics is a booming field with application in many areas of medicine, including dentistry. Nevertheless, proteomic characterization of bone tissues in oral surgery is not still commonly used. The main reason is involvement of demanding analytical approaches due to insoluble chatacter of bone tissues. The goal of this work was to develop and apply straightforward methodology that could lead to the routine use of proteomics in this area as well. Using porcine jawbones as a model samples, a technique was developed allowing identifying about hundreds of proteins thanks to their trypsin digestion directly in bone tissues ("in-bone digestion") followed by analysis using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). This technique was subsequently applied to the analysis of human maxillary and mandibular bone tissues obtained during surgical procedures. In both maxillary and mandibular bone samples, it was possible to identify a considerable amount of proteins using the "in-bone digestion" technique. Additionaly, the mathematical analysis of the obtained data was able to distinguish between the inflammatory and healthy tissues. The approach based on direct cleavage was subsequently successfully extended to the analysis of in vitro models of human bone tissues. Direct...

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